IGEM:IMPERIAL/2009/201009

Dear all, when editing please use comment with name under each section. Feedback will be greatly appreciated. DK & JF

THE PRESENTATION SCRIPT

30 Slides:
2 = title page, team
5 = intro
15 = platform, modules, results etc
5 = applications
3 = summary, ethics, achievements

Slide 1 (Title slide)

• Include: E.ncapsulator text, imperial logo, encapsulator logo
• Hold up pills in tube JF & DK:
o ‘This pill contains x million cells’
o ‘Each cell contains x amount of protein’
o ‘With my pill, I can treat the genetic disease Phenylketonuria’
o ‘And mine is 6 times as powerful as morphine…’

• ‘Welcome to Imperial’s 2009 project: The Encapsulator’.

Slide 2 (Team slide)

• Include: Team picture, advisor pictures, student disciplines (2 Biologists, 2 Biochemists, 4 Bioengineers)
• The 2009 Imperial Team consists of an equal mix of Engineers and Life scientists and of course a number of advisors.

Slide 3 (Problem slide)

o Introduce the problem - why are we doing this project?!
o Theoretical advantages: Diversity, Bioactivity, Biosynthesis
o Projected annual sales in 2010 = $52.2 billion
o Bottleneck: Oral availability
o Use example to illustrate: Insulin discovered over 80 years ago but still using injections.

Royah Vaezi - Would like to know more about the first bullet point, e.g. the actual script.

Slide 4 (Problem slide 2 - problem drilled down)

• (Get an equivalent to the computer transistor diagram)
o All orally delivered polypeptides share a common predicament: the stomach.
o The stomach is a protease rich acid bath which serves to cut polypeptides into small, non functional pieces.
o If you can bypass the stomach, this will dramatically increase oral availability.
o While protein engineering requires precise knowledge can be used to confer acid resistance, it requires a reductionist understanding of an individual polypeptide. Decoupling this problem from the protein facilitates the possibility of a universal solution.
o certain polypeptides can be modified to enhance acid resistance, it requires an indepth knowledge of the physical
o However, the Imperial team have come up with a solution. The E.ncapsulator is a versatile manufacturing and delivery platform by which therapeutics can ferried through the stomach.
o By decoupling the physical structure of the protein from the problem,

Slide 5 (Mission statement - so they know exactly what we are doing)

• Manufacture a pill with the scope to contain any polypeptide
• And successfully delivery this polypeptide past the stomach.

Slide 6 (Specs)

• Polypeptide Production
o Specifications: To produce any polypeptide.
o Solution: Proteins can be genetically encoded - logical to use biosynthesis over chemical synthesis
• Polypeptide Protection
o Specification: A) Against stomach acid of pH 1-2 and released past the stomach. Non toxic.
o B) Against dehydration
o Solution: A) Acid resistant capsule - also means we don’t have to purify protein product. Based on natural sources of acid resistance, Lactobacillus, E.coli and B.subtilis were shortlisted as potential chassis. E.coli was deemed the most suitable - possesses a broad range of acid resistance strategies.
o B) Synthesis of a preservative in addition keeping the polypeptide inside the cell is increasingly stable.
• Be safe & socially acceptable
o Specification: A) No risks B) Inanimate
o Solution: Genome deletion - doesn’t damage protein or membrane. Using k12 strain as safe.
• Dosage Control
o Specification: Tunable dosage & robust quality control
o Solution: Macroencapsulation of cells

Slide 7 (Solution Overview Picture)

• Here’s an overview of The Encapsulator system.
• Our E.coli chassis progresses through a series of defined modules culminating in the production of a safe, inanimate pill. This sequential process involves M1, polypeptide production, M2, protective encapsulation and M3, genome deletion.
• Taking this modular approach allows for specialisation (bacteria can devote all it’s energy to the specific task required for that module - increases efficiency).
• However, we’ve taken this 1 step further to make it compatible for industrial production. Integration modules have been designed and incorporated, adding a means of control to this platform.

Royah Vaezi - What are the advantages of the integration modules over a genetic timer? Why was this approach taken? I think this needs to be emphasised as it is the key SynBio aspect of our project.

Slide 8 (Temporal control specs)

- Justify why we use these
• Module 1 specs: Grow up the bacteria as protein production is metabolically demanding and therefore need enough bacteria to carry out the task.
• Solution: Chemoinduction is proven technology and is currently used in industry. The output is instantaneous, long lasting and either on or off at any one time.
• M2 specs: Tuneable delay, synchronous response at a population level.
• Solution: Media defined autoinduction switch - we didn’t use a timer as they are unable to provide a long enough delay, they can fall out of phase and require multiple genes
• M3 specs: Has to be tight (restriction enzymes), responsive to physical cues
• Solution: Thermoinduction, signal can pass through capsule.

Slide 9 (Entire Solution Overview)

• So here’s the final system. We will now tell you the story of our Encapsulator…

Royah Vaezi - It's certainly coming together. Just watch out on timing as you go through. You don't want to spend too much time at the beginning and have to rush through at the end. Also remember think about the speed at which you'll be talking as this will determine how much ou can say. It may be better to cut down on jargon and still get the point across clearly and concisely, rather than using too many words and running out of time for other things. I'll keep an eye out for further developments. Keep up the good work guys!